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Development and Evaluation of Detection Method for Pathogenic Bacteria from...
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Development and Evaluation of Detection Method for Pathogenic Bacteria from Food

A special issue of Pathogens (ISSN 2076-0817). This special issue belongs to the section "Bacterial Pathogens".

Deadline for manuscript submissions: closed (31 August 2024) | Viewed by 15558

Special Issue Editors

Dr. Luca Nalbone

E-Mail Website
Guest Editor
Department of Veterinary Science, University of Study of Messina, 98168 Messina, Italy
Interests: food microbiology; foodborne pathogens; antimicrobial resistance; antimicrobial tolerance; antimicrobial persistence
Special Issues, Collections and Topics in MDPI journals
Dr. Filippo Giarratana

E-Mail Website
Guest Editor
Department of Veterinary Science, University of Study of Messina, 98168 Messina, Italy
Interests: food microbiology; foodborne disease; predictive microbiology; natural antimicrobials; food risk assessment
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Nowadays, increasing public attention is paid to food safety which is jeopardized by new global challenges. The growth of the world population and the consequent demand for food will lead to a significant increase in food production which will require extra efforts to ensure high standards of quality and safety. The scientific community is therefore called to find new solutions to ensure adequate control of foods and guarantee their safety. Over the years, microbial detection techniques have evolved frequently compared to conventional methods and, even today, always new and alternative techniques are proposed. The development of increasingly rapid and effective techniques for the detection of pathogenic microorganisms in foods is crucial to guarantee food safety in view of the significant increase in production in the coming next years.

Both original research and review articles are welcomed. Potential topics include, but are not limited to:

  • New techniques for the detection and enumeration of microorganisms in food;
  • Validation of techniques for the detection and enumeration of microorganisms in food;
  • Evaluation and comparison of the effectiveness of conventional techniques for the detection and enumeration of microorganisms in food;

Dr. Luca Nalbone
Dr. Filippo Giarratana
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Pathogens is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2200 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • food microbiology
  • detection and enumeration of microorganisms in food
  • microbial detection techniques
  • foodborne bacteria
  • food safety

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Published Papers (7 papers)

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Research

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13 pages, 817 KiB  
Article
Salmonella Infantis Adhesion to Various Surfaces and In Vitro Antimicrobial Efficacy of Commercial Disinfectants
by Katja Kranjc, Jana Avberšek, Neva Šemrov, Olga Zorman-Rojs and Darja Barlič-Maganja
Pathogens 2024, 13(11), 999; https://doi.org/10.3390/pathogens13110999 - 14 Nov 2024
Cited by 1 | Viewed by 947
Abstract
Salmonella Infantis poses a significant challenge in poultry production due to its persistence and resistance to disinfectants. This study investigated the survival of the S. Infantis strain on different surfaces and evaluated the efficacy of disinfectants in both preventing and treating biofilms. The [...] Read more.
Salmonella Infantis poses a significant challenge in poultry production due to its persistence and resistance to disinfectants. This study investigated the survival of the S. Infantis strain on different surfaces and evaluated the efficacy of disinfectants in both preventing and treating biofilms. The survival of the tested S. Infantis strain was assessed on plastic and stainless steel surfaces after 24 and 48 h. The minimum inhibitory concentrations (MICs) of five disinfectants were determined, and their antiadhesion effectiveness was evaluated using crystal violet. The efficacy of biofilm treatment was evaluated by cell culturability. The results showed that the adhesion of S. Infantis was significantly higher on the plastic surface. The disinfectants were effective at reducing biofilm formation only within the first 24 h. Fresh solutions of disinfectants based on quaternary ammonium compounds exhibited the highest antimicrobial efficacy, while chlorocresol was the most effective for both the prevention and treatment of biofilms. The study results suggest that the presence of plastic surfaces may contribute to the dissemination of Salmonella. Additionally, the effectiveness of disinfectants varied based on storage conditions and contact time, while biofilms demonstrated reduced susceptibility compared to planktonic cells. However, given the laboratory scale of this study, further validation on a commercial scale is necessary to confirm these findings. Full article
(This article belongs to the Special Issue Development and Evaluation of Detection Method for Pathogenic Bacteria from Food)
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7 pages, 860 KiB  
Article
Innovative Multiplex PCR Assay for Detection of tlh, trh, and tdh Genes in Vibrio parahaemolyticus with Reference to the U.S. FDA’s Bacteriological Analytical Manual (BAM)
by Seong Bin Park and Yan Zhang
Pathogens 2024, 13(9), 774; https://doi.org/10.3390/pathogens13090774 - 7 Sep 2024
Cited by 2 | Viewed by 1713
Abstract
Vibrio parahaemolyticus is an important foodborne bacterium that causes severe gastroenteritis following the consumption of contaminated seafood. To identify V. parahaemolyticus and determine its pathogenicity, the U.S. Food and Drug Administration (FDA)’s Bacteriological Analytical Manual (BAM) recommends a multiplex polymerase chain [...] Read more.
Vibrio parahaemolyticus is an important foodborne bacterium that causes severe gastroenteritis following the consumption of contaminated seafood. To identify V. parahaemolyticus and determine its pathogenicity, the U.S. Food and Drug Administration (FDA)’s Bacteriological Analytical Manual (BAM) recommends a multiplex polymerase chain reaction (PCR) protocol to simultaneously detect the species-specific thermolabile hemolysin (tlh) gene and the pathogenic thermostable-related hemolysin (trh) and thermostable-direct hemolysin (tdh) genes. However, this assay has shown two limitations: difficulty in separating the amplicons of the trh (486 bp) and tlh (450 bp) genes due to their highly similar sizes, and the weaker band exhibited by the tdh gene amplicon (270 bp). The present study aimed to improve the BAM’s multiplex PCR assay by separating the three amplicons with similar intensity. A new primer set was applied for the tlh gene (369 bp) alongside the existing primers for the trh and tdh genes. The amplicons for the three genes were effectively separated by electrophoresis on a 2% tris-borate-EDTA (TBE) agarose gel within 45 min. Primer concentrations of 0.25 µM for three genes produced a significant amount of amplicons among various combinations of primer concentrations with 35 PCR cycles. This assay exhibited a detection limit of 10 pg of bacterial DNA, demonstrating its high sensitivity. It did not display amplicons from nine Vibrio species known to be human pathogens or from 18 well-documented foodborne pathogens. Therefore, the present multiplex PCR protocol could help overcome the limitations of existing assays and provide a more reliable method for detecting the three genes of V. parahaemolyticus. Full article
(This article belongs to the Special Issue Development and Evaluation of Detection Method for Pathogenic Bacteria from Food)
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17 pages, 475 KiB  
Article
Use of Matrix-Assisted and Laser Desorption/Ionization Time-of-Flight Technology in the Identification of Aeromonas Strains Isolated from Retail Sushi and Sashimi
by Luca Nalbone, Salvatore Forgia, Federico Pirrone, Filippo Giarratana and Antonio Panebianco
Pathogens 2024, 13(6), 432; https://doi.org/10.3390/pathogens13060432 - 21 May 2024
Viewed by 1104
Abstract
The genus Aeromonas includes well-known pathogenic species for fishes and humans that are widely distributed in the aquatic environment and foods. Nowadays, one of the main issues related to wild Aeromonas isolates is their identification at the species level, which is challenging using [...] Read more.
The genus Aeromonas includes well-known pathogenic species for fishes and humans that are widely distributed in the aquatic environment and foods. Nowadays, one of the main issues related to wild Aeromonas isolates is their identification at the species level, which is challenging using classical microbiological and biomolecular methods. This study aims to test MALDI-TOF MS technology in the identification of Aeromonas strains isolated from n. 60 retail sushi and sashimi boxes using an implemented version of the SARAMIS software V4.12. A total of 43 certified Aeromonas strains were used to implement the SARAMIS database by importing the spectra obtained from their identification. The original SARAMIS version (V4.12) failed to recognize 62.79% of the certified strains, while the herein-implemented version (V4.12plus) allowed the identification of all the certified strains at least to the genus level with a match of no less than 85%. Regarding the sushi and sashimi samples, Aeromonas spp. was detected in n. 18 (30%) boxes. A total of 127 colonies were identified at the species level, with A. salmonicida detected as the most prevalent species, followed by A. bestiarum and A. caviae. Based on the results of the present study, we could speculate that MALDI-TOF technology could be a useful tool both for the food industry to monitor product contamination and for clinical purposes to make diagnoses effectively and quickly. Full article
(This article belongs to the Special Issue Development and Evaluation of Detection Method for Pathogenic Bacteria from Food)
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8 pages, 248 KiB  
Communication
Verification of a Rapid Analytical Method for the Qualitative Detection of Listeria spp. and Listeria monocytogenes by a Real-Time PCR Assay according to EN UNI ISO 16140-3:2021
by Veronica Bolzon, Michela Bulfoni, Massimo Pesando, Alessandro Nencioni and Emanuele Nencioni
Pathogens 2024, 13(2), 141; https://doi.org/10.3390/pathogens13020141 - 4 Feb 2024
Cited by 3 | Viewed by 2572
Abstract
Microbial contamination and foodborne infections are a significant global public health concern. For this reason, the detection, monitoring, and characterization of pathogens represent a significant challenge in quality control settings. Standard approaches, such as culture methods and biochemical tests, are known to be [...] Read more.
Microbial contamination and foodborne infections are a significant global public health concern. For this reason, the detection, monitoring, and characterization of pathogens represent a significant challenge in quality control settings. Standard approaches, such as culture methods and biochemical tests, are known to be very time-consuming and intensive. Conversely, molecular technologies based on the genomic identification of bacteria are quick and low-cost. Listeria monocytogenes is an opportunistic pathogen and a major concern especially in food industries. It is important to understand and implement multiple quality control measures to control Listeria infection risk and prevent the contamination of products. Standardized detection and confirmation tests such as the API Listeria test, MALDI-TOF MS, and PCR analysis are available. The aim of our work is to provide a specific molecular method, designed according to the EN UNI ISO 16140-3:2021, for the specific detection, monitoring, and characterization of Listeria spp. and Listeria monocytogenes contamination. The verification of this new rapid approach by real-time PCR (qPCR) overcomes the limitations of culture-based techniques, meeting all the verification criteria required by ISO guidelines, including implementation and item confirmation. This system offers a powerful approach to the real-time assessment of food safety, useful for industry self-monitoring and regulatory inspection. Full article
(This article belongs to the Special Issue Development and Evaluation of Detection Method for Pathogenic Bacteria from Food)
13 pages, 2874 KiB  
Article
Development of Multienzyme Isothermal Rapid Amplification (MIRA) Combined with Lateral-Flow Dipstick (LFD) Assay to Detect Species-Specific tlh and Pathogenic trh and tdh Genes of Vibrio parahaemolyticus
by Seong Bin Park and Yan Zhang
Pathogens 2024, 13(1), 57; https://doi.org/10.3390/pathogens13010057 - 6 Jan 2024
Cited by 6 | Viewed by 2587
Abstract
Vibrio parahaemolyticus causes severe gastroenteritis in humans after consuming contaminated raw or undercooked seafood. A species-specific marker, the thermolabile hemolysin (tlh) gene, and two pathogenic markers, thermostable-related hemolysin (trh) and thermostable-direct hemolysin (tdh) genes, have been used [...] Read more.
Vibrio parahaemolyticus causes severe gastroenteritis in humans after consuming contaminated raw or undercooked seafood. A species-specific marker, the thermolabile hemolysin (tlh) gene, and two pathogenic markers, thermostable-related hemolysin (trh) and thermostable-direct hemolysin (tdh) genes, have been used to identify V. parahaemolyticus and determine its pathogenicity using both PCR and qPCR assays. To enable testing in field conditions with limited resources, this study aimed to develop a simple and rapid method to detect the species-specific (tlh) and pathogenic (trh and tdh) genes of V. parahaemolyticus using multienzyme isothermal rapid amplification (MIRA) combined with a lateral-flow dipstick (LFD). The amplification of the tlh, trh, and tdh genes could be completed within 20 min at temperatures ranging from 30 to 45 °C (p < 0.05). The test yielded positive results for V. parahaemolyticus but produced negative results for nine Vibrio species and eighteen foodborne pathogenic bacterial species. MIRA-LFD could detect 10 fg of DNA and 2 colony-forming units (CFU) of V. parahaemolyticus per reaction, demonstrating a sensitivity level comparable to that of qPCR, which can detect 10 fg of DNA and 2 CFU per reaction. Both MIRA-LFD and qPCR detected seven tlh-positive results from thirty-six oyster samples, whereas one positive result was obtained using the PCR assay. No positive results for the trh and tdh genes were obtained from any oyster samples using MIRA-LFD, PCR, and qPCR. This study suggests that MIRA-LFD is a simple and rapid method to detect species-specific and pathogenic genes of V. parahaemolyticus with high sensitivity. Full article
(This article belongs to the Special Issue Development and Evaluation of Detection Method for Pathogenic Bacteria from Food)
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20 pages, 11145 KiB  
Article
Addressing Current Challenges in Poultry Meat Safety: Development of a Cultivation and Colony Hybridization Approach to Recover Enterotoxigenic Clostridium perfringens from Broiler Chicken Carcasses
by Rosette Kakese Mukosa, Alexandre Thibodeau, John Morris Fairbrother, William Thériault and Marie-Lou Gaucher
Pathogens 2024, 13(1), 30; https://doi.org/10.3390/pathogens13010030 - 28 Dec 2023
Cited by 1 | Viewed by 1869
Abstract
Enterotoxigenic Clostridium perfringens is one of the main causes of foodborne illness in Canada. The use of a conventional bacterial culture approach to isolate enterotoxigenic C. perfringens from poultry meat is common. This approach is based on the phenotype attributable to a double [...] Read more.
Enterotoxigenic Clostridium perfringens is one of the main causes of foodborne illness in Canada. The use of a conventional bacterial culture approach to isolate enterotoxigenic C. perfringens from poultry meat is common. This approach is based on the phenotype attributable to a double hemolysis phenomenon, whereas few enterotoxigenic strains of C. perfringens produce it, which further complicates the study of the reservoirs of this important pathogen. The objectives of the current study were to validate the ability of a digoxigenin-labeled probe to detect the C. perfringens cpe gene and to validate the use of either a filtration or a direct plating approach, combined with colony hybridization to detect enterotoxigenic C. perfringens. Pure DNA and pure colonies of enterotoxigenic C. perfringens and broiler chicken carcass rinsate samples were subjected to colony hybridization. The results showed that the synthesized DNA probe can detect the cpe gene from both DNA and pure colonies of enterotoxigenic C. perfringens, and from colonies grown from carcass rinsates artificially contaminated with enterotoxigenic C. perfringens. Our study suggests that this isolation method is a promising tool for a better understanding of the epidemiology of this zoonotic pathogen. Full article
(This article belongs to the Special Issue Development and Evaluation of Detection Method for Pathogenic Bacteria from Food)
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Review

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18 pages, 2264 KiB  
Review
Advancements in Detection Methods for Salmonella in Food: A Comprehensive Review
by Aayushi Patel, Andrew Wolfram and Taseen S. Desin
Pathogens 2024, 13(12), 1075; https://doi.org/10.3390/pathogens13121075 - 7 Dec 2024
Viewed by 3644
Abstract
Non-typhoidal Salmonella species are one of the leading causes of gastrointestinal disease in North America, leading to a significant burden on the healthcare system resulting in a huge economic impact. Consequently, early detection of Salmonella species in the food supply, in accordance with [...] Read more.
Non-typhoidal Salmonella species are one of the leading causes of gastrointestinal disease in North America, leading to a significant burden on the healthcare system resulting in a huge economic impact. Consequently, early detection of Salmonella species in the food supply, in accordance with food safety regulations, is crucial for protecting public health, preventing outbreaks, and avoiding serious economic losses. A variety of techniques have been employed to detect the presence of this pathogen in the food supply, including culture-based, immunological, and molecular methods. The present review summarizes these methods and highlights recent updates on promising emerging technologies, including aptasensors, Surface Plasmon Resonance (SPR), and Surface Enhanced Raman Spectroscopy (SERS). Full article
(This article belongs to the Special Issue Development and Evaluation of Detection Method for Pathogenic Bacteria from Food)
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